Human Mesenchymal Stem Cell Verification Flow Kit

Name: Human Mesenchymal Stem Cell Verification Flow Kit
Brand: R&D Systems
SKU: FMC020
Price: 1360 USD

Catalog # FMC020 | R&D Systems, Inc. a Bio-Techne Brand

Citations (10)

Product Datasheet / COA / SDS

Key Product Details

Setting the standard in quality research reagents

Summary for Human Mesenchymal Stem Cell Verification Flow Kit

Kit Summary

Antibodies for MSC verification according to the markers proposed by the International Society for Cellular Therapy.

Why use the ISCT-recommended Markers to Verify MSC Identity?

The different methods used to isolate, culture, differentiate, and define mesenchymal stem cells (MSCs) can lead to experimental variability and contradictory data. In an attempt to standardize the definition of mesenchymal stem cells, in 2006, the International Society for Cellular Therapy (ISCT) proposed the minimal criteria to define MSCs.¹ See Details

Included in these criteria were recommendations for the positive and negative expression of a specific set of surface antigens. Specifically, the ISCT criteria recommends that 95% or greater of the MSC population must express CD73/5'-Nucleotidase, CD90/Thy1, and CD105/Endoglin as measured by flow cytometry and that 2% or less of the MSC population should express CD34, CD45, CD11b/Integrin alpha M or CD14, CD79 alpha or CD19, and HLA Class II.

R&D Systems Human Mesenchymal Stem Cell Marker Verification Multi-Color Flow Cytometry Kit (Catalog # FMC020) includes all of the antibodies needed to assess MSC marker expression according to the ISCT's definition of human MSCs. To fulfill both the phenotypic and functional definition of human MSCs according to the ISCT criteria, cells can be assessed using both the Human Mesenchymal Stem Cell Marker Verification Multi-Color Flow Cytometry kit and the Human Mesenchymal Stem Cell Functional Identification Kit.

References

Dominici, M. et al. (2006) Cytotherapy 8:315.

Human Mesenchymal Stem Cell Marker Verification Multi-Color Flow Cytometry Kit:

Provides antibodies to the MSC markers recommended by the ISCT's minimal criteria to define human MSCs.
Can be used with the Human MSC Functional ID kit to define MSCs according to the ISCT recommendations.
Reduces experimental variability by defining the starting cell population.

Mesenchymal Stromal Cells or Mesenchymal Stem Cells?

The term ‘mesenchymal stromal cells’ is commonly used to describe a heterogeneous population of cultured cells that are adherent to plastic, have a distinct morphology, and express a specific set of marker proteins. Within this heterogeneous population are cells referred to as ‘mesenchymal stem cells.’ See Details

Mesenchymal stem cells are multipotent, self-renewing cells that have the ability to differentiate into adipocytes, chondrocytes, and osteoblasts when cultured in vitro.

2006 Proposed Change to MSC Nomenclature

Although mesenchymal stromal cells were once referred to as ‘mesenchymal stem cells,' a change to ‘mesenchymal stromal cells' was proposed by the International Society for Cellular Therapy in 2006.¹

The change in nomenclature originates from two important factors:

Methods used to isolate mesenchymal stem cells yield a heterogeneous population of cells with only a fraction of these cells demonstrating multipotency.
The absence of direct evidence that mesenchymal stem cells can self-renew and differentiate in vivo.

Use of Mesenchymal Stem and Stromal Cell Terminology

Data supporting MSC self-renewal and multipotency have been obtained using in vitro conditions, which does not adequately reflect the in vivo environment. The lack of in vivo data has led some researchers to question the validity of the term ‘mesenchymal stem cell' providing further support for the use of ‘mesenchymal stromal cells' to describe MSCs.² While ‘mesenchymal stromal cells' may be the more scientifically accurate term for MSCs, the two terms are often used interchangeably in the literature. R&D Systems recognizes the use of both mesenchymal stem cells and mesenchymal stromal cells and uses ‘MSC' to indicate mesenchymal stem/stromal cells to account for both designations.

Definitions of Mesenchymal Stromal Cells and Mesenchymal Stem Cells

Mesenchymal Stromal Cells – A heterogeneous population of cultured cells with similar characteristics such as the ability to adhere to plastic and the expression of specific marker proteins.
Mesenchymal Stem Cells – A subpopulation of mesenchymal stromal cells that have the capacity to self-renew and differentiate into mesodermal lineages when cultured in vitro. The capacity to self-renew and differentiate in vivo has yet to be clearly demonstrated for mesenchymal stem cells.

References

Dominici, M. et al. (2006) Cytotherapy 8:315.
Keating, A. (2012) Cell Stem Cell 10:709.

Kit Components

Positive Markers:

CD90-APC Mouse IgG_2A; Clone Thy-1A1; 500 μL
CD73-CFS Mouse IgG_2B; Clone 606112; 500 μL
CD105-PerCP Mouse IgG₁; Clone 166707; 500 μL

Negative Markers:

Negative Marker Cocktail (500 μL) that includes:
- CD45-PE Mouse IgG₁; Clone 2D1
- CD34-PE Mouse IgG₁; Clone QBEnd10
- CD11b-PE Mouse IgG_2B; Clone 238446
- CD79A-PE Mouse IgG₁; Clone 706931
- HLA-DR-PE Mouse IgG₁; Clone L203

Isotype Controls:

Mouse IgG_2A-APC Isotype Control; Clone 20102; 500 μL
Mouse IgG_2B-CFS Isotype Control; Clone 133303; 500 μL
Mouse IgG₁-PerCP Isotype Control; Clone 11711; 500 μL
Negative Isotype Cocktail (500 μL) that includes:
- Mouse IgG₁-PE Isotype Control; Clone 11711
- Mouse IgG_2B-PE Isotype Control; Clone 133303
125 mL of 1X Staining Buffer

Stability and Storage

Store at 2 °C to 8 °C in the dark. Use within 6 months of receipt.

The term 'mesenchymal stem cells' (MSCs) is most commonly used to describe multipotent self-renewing cells that can be differentiated in vitro to generate adipocytes, chondrocytes, and osteoblasts. However, because these biological properties and hierarchical relationships remain to be clearly demonstrated in vivo, the term 'multipotent mesenchymal stromal cells' is often used to distinguish cultured cells from their in vivo precursors. Originally discovered in mouse bone marrow, multipotent mesenchymal stromal cells cultured from a variety of species and tissue types, have been shown to differentiate into progeny of additional lineages including, cardiomyocytes, endothelial cells, hepatocytes, and neural cells. Again, the physiological relevance of these findings remains to be determined.

Product Specifications for Human Mesenchymal Stem Cell Verification Flow Kit

Species

Human

Scientific Data Examples for Human Mesenchymal Stem Cell Verification Flow Kit

	Human Bone-Marrow Derived Cells Fulfill the ISCT's Definition of Human MSCs Based on Marker Expression. Human bone marrow-derived mesenchymal stem cells were stained using the antibodies and reagents provided in the Human Mesenchymal Stem Cell Marker Verification Multi-Color Flow Cytometry Kit (Catalog # FMC020). The data shows positive expression of MSC-associated surface antigens CD73, CD90, and CD105. In contrast, minimal expression of antigens recognized by the Negative Marker Cocktail was detected.
	Surface Antigens Expressed by KG-1 Acute Myelogenous Leukemia Cells and Peripheral Blood Mononuclear Cells are Recognized by the Antibodies Included in the Negative Marker Cocktail. KG-1 acute myelogenous leukemia cells (A) and peripheral blood mononuclear cells (PBMCs) (B) were stained using the Negative Marker Cocktail included in the Human Mesenchymal Stem Cell Marker Verification Multi-Color Flow Cytometry Kit (Catalog # FMC020). The cells demonstrate positive expression of antigens recognized by the antibodies included in the Negative Marker Cocktail (filled histogram) over the Negative Isotype Control Cocktail (open histograms).
	Human MSCs Fulfill the ISCT’s definition of Human Mesenchymal Stem Cells Based on Functional Differentiation. Human mesenchymal stem cells were cultured in StemXVivo™ Mesenchymal Stem Cell Expansion Media (Catalog # CCM004) and differentiation was induced as indicated using the media supplements included in the Human Mesenchymal Stem Cell Functional Identification Kit (Catalog # SC006). The kit also contains a Goat Anti-Mouse FABP-4 Antigen Affinity-purified Polyclonal Antibody (adipocytes), a Goat Anti-Human Aggrecan Antigen Affinity-purified Polyclonal Antibody (chondrocytes), and a Mouse Anti-Human Osteocalcin Monoclonal Antibody (osteocytes) for the confirmation of differentiation status. The cells were stained using the NorthernLights™ 557-conjugated Donkey Anti-Goat (Catalog # NL001; red) or Anti-Mouse (Catalog # NL007; red) IgG Secondary Antibodies, and the nuclei were counterstained with DAPI (blue).

Preparation & Storage

Shipping Conditions	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage	Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, mesenchymal stem cells can be analyzed for marker expression using the following procedure:

Incubate the cells with the provided antibodies
Wash the cells
Resuspend in Flow Cytometry Staining Buffer
Analyze by flow cytometry

View Graphic Procedure

Reagents Provided

Kit Components

Reagents supplied in the Human Mesenchymal Stem Cell Marker Verification Multi-Color Flow Cytometry Kit (Catalog # FMC020):

Positive Markers:

CD90-APC Mouse IgG_2A; Clone Thy-1A1; 500 μL
CD73-CFS Mouse IgG_2B; Clone 606112; 500 μL
CD105-PerCP Mouse IgG₁; Clone 166707; 500 μL

Negative Markers:

Negative Marker Cocktail (500 mL) that includes:
- CD45-PE Mouse IgG₁; Clone 2D1
- CD34-PE Mouse IgG₁; Clone QBEnd10
- CD11b-PE Mouse IgG_2B; Clone 238446
- CD79A-PE Mouse IgG₁; Clone 706931
- HLA-DR-PE Mouse IgG₁; Clone L203

Isotype Controls:

Mouse IgG_2A-APC Isotype Control; Clone 20102; 500 μL
Mouse IgG_2B-CFS Isotype Control; Clone 133303; 500 μL
Mouse IgG₁-PerCP Isotype Control; Clone 11711; 500 μL
Negative Isotype Cocktail (500 μL) that includes:
- CD45-PE Mouse IgG₁; Clone 2D1
- CD34-PE Mouse IgG₁; Clone QBEnd10
- CD11b-PE Mouse IgG_2B; Clone 238446
- CD79A-PE Mouse IgG₁; Clone 706931
- HLA-DR-PE Mouse IgG₁; Clone L203
125 mL of 1X Staining Buffer

Other Supplies Required

Reagents

Fc receptor blocking reagents (optional)

Materials

Flow Cytometry/FACS™ Tubes (5 mL round-bottom polystyrene tubes)
Pipette Tips and Pipettes

Equipment

Clinical centrifuge
Hemocytometer
Flow Cytometer

Procedure Overview

Wash cells with 2 mL of Staining Buffer.

Perform a cell count.

Add Fc receptor blocking reagents (optional).
Transfer 1 x 10⁵ cells in 100 μL into a 5 mL Flow Cytometry tube.

Add 10 μL of each positive antibody and 10 μL of the Negative Marker Cocktail or add 10 μL of each positive isotype control antibody and 10 μL of the Negative Isotype Control Cocktail.
Incubate the samples for 30-45 minutes at room temperature in the dark.

add positive antibody and positive isotype control antibody

Wash the cells with Staining Buffer.
Resuspend cell pellets in 200-400 μL of Staining Buffer.

Analyze the cells for marker expression by flow cytometry.

Analyze the cells for marker expression by flow cytometry.

Citations for Human Mesenchymal Stem Cell Verification Flow Kit (10)

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Citations are publications that use Bio-Techne products. Selected citations for Human Mesenchymal Stem Cell Verification Flow Kit include:

Species: Human
Sample Types: Whole Cells

Yaylac? et al. (2023-05-17), An enzyme-free technique enables the isolation of a large number of adipose-derived stem cells at the bedside Scientific reports
PMID: 37198228
Species: Human
Sample Types: Whole Cells

AJ Janvier et al. (2022-10-31), Multimodal analysis of the differential effects of cyclic strain on collagen isoform composition, fibril architecture and biomechanics of tissue engineered tendon Journal of tissue engineering, 2022-113(0):2041731422113.
PMID: 36339372
DM Iacomi et al. (2022-08-10), Generation of an Immortalized Human Adipose-Derived Mesenchymal Stromal Cell Line Suitable for Wound Healing Therapy International Journal of Molecular Sciences, 2022-023(16).
PMID: 36012192
Species: Human
Sample Types: Whole Cells

B Sikora et al. (2021-10-07), Paracrine activity of adipose derived stem cells on limbal epithelial stem cells Scientific Reports, 2021-111(1):19956.
PMID: 34620960
Species: Human
Sample Types: Whole Cells

M Yoshimatsu et al. (2021-02-11), In vivo regeneration of rat laryngeal cartilage with mesenchymal stem cells derived from human induced pluripotent stem cells via neural crest cells Stem Cell Research, 2021-052(0):102233.
PMID: 33607469
Species: Human
Sample Types: Whole Cells

I Pountos et al. (2021-01-10), NSAIDs inhibit bone healing through the downregulation of TGF-&beta3 expression during endochondral ossification Injury, 2021-00(0).
PMID: 33472741
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry

S Mount et al. (2019-11-04), Physiologic expansion of human heart-derived cells enhances therapeutic repair of injured myocardium Stem Cell Res Ther, 2019-110(1):316.
PMID: 31685023
Species: Human
Sample Types: Whole Cells

B Sikora et al. (2019-10-24), Adipose-derived stem cells undergo differentiation after co-culture with porcine limbal epithelial stem cells Stem Cell Res, 2019-141(0):101609.
PMID: 31706096
Species: Human
Sample Types: Whole Cells

JM McKinney et al. (2019-01-29), Therapeutic efficacy of intra-articular delivery of encapsulated human mesenchymal stem cells on early stage osteoarthritis Eur Cell Mater, 2019-037(0):42-59.
PMID: 30693466
Species: Human
Sample Types: Whole Cells

A Skubis et al. (2017-11-24), Impact of Antibiotics on the Proliferation and Differentiation of Human Adipose-Derived Mesenchymal Stem Cells Int J Mol Sci, 2017-118(12).
PMID: 29186789

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Product Documents for Human Mesenchymal Stem Cell Verification Flow Kit

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Human Mesenchymal Stem Cell Verification Flow Kit

Catalog # FMC020 | R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

Summary for Human Mesenchymal Stem Cell Verification Flow Kit

Antibodies for MSC verification according to the markers proposed by the International Society for Cellular Therapy.

Why use the ISCT-recommended Markers to Verify MSC Identity?

References

Human Mesenchymal Stem Cell Marker Verification Multi-Color Flow Cytometry Kit:

Mesenchymal Stromal Cells or Mesenchymal Stem Cells?

2006 Proposed Change to MSC Nomenclature

Use of Mesenchymal Stem and Stromal Cell Terminology

Definitions of Mesenchymal Stromal Cells and Mesenchymal Stem Cells

References

Kit Components

Positive Markers:

Negative Markers:

Isotype Controls:

Stability and Storage

Product Specifications for Human Mesenchymal Stem Cell Verification Flow Kit

Scientific Data Examples for Human Mesenchymal Stem Cell Verification Flow Kit

Preparation & Storage

Assay Procedure

Briefly, mesenchymal stem cells can be analyzed for marker expression using the following procedure:

Kit Components

Reagents supplied in the Human Mesenchymal Stem Cell Marker Verification Multi-Color Flow Cytometry Kit (Catalog # FMC020):

Positive Markers:

Negative Markers:

Isotype Controls:

Other Supplies Required

Reagents

Materials

Equipment

Procedure Overview

Citations for Human Mesenchymal Stem Cell Verification Flow Kit (10)

Product Documents for Human Mesenchymal Stem Cell Verification Flow Kit

Product Datasheets

COA

Certificate of Analysis

SDS

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